Downregulation of microRNA-144 inhibits proliferation and promotes the apoptosis of myelodysplastic syndrome cells through the activation of the AKAP12-dependent ERK1/2 signaling pathway

Cell Signal. 2020 Apr:68:109493. doi: 10.1016/j.cellsig.2019.109493. Epub 2019 Dec 3.

Abstract

Background: Myelodysplastic syndromes (MDS) represent a family of hematopoietic stem cell disorders characterized by ineffective hematopoiesis. While the functions of many microRNAs have been identified in MDS, microRNA-144 (miR-144) remains poorly understood. Thus, the aim of the present study was to determine the effects of miR-144 on cell proliferation and apoptosis in MDS cells and mechanism thereof.

Methods: MDS-related microarrays were used for screening differentially expressed genes in MDS. The relationship between miR-144 and A-kinase anchoring protein 12 (AKAP12) was determined by a dual luciferase reporter gene assay. Subsequently, gain- and loss-function approaches were used to assess the effects of miR-144 and AKAP12 on cell proliferation, cell cycle and cell apoptosis by MTT assay and flow cytometry. Following the induction of a mouse model with MDS, the tumor tissues were extract for evaluation of apoptosis and the expression of miR-144, AKAP12, and the relevant genes associated with extracellular-regulated protein kinases 1/2 (ERK1/2) signaling pathway and apoptosis.

Results: We observed significantly diminished expression of AKAP12 in MDS samples. miR-144 directly bound to AKAP12 3'UTR and reduced its expression in hematopoietic cells. Downregulation of miR-144 or upregulation of AKAP12 was observed to prolong cell cycle, inhibit cell proliferation, and induce apoptosis, accompanied by increased expression of AKAP12, p-ERK1/2, caspase-3, caspase-9, Bax, and p53, as well as decreased expression of Bcl-2. The transplanted tumors in mice with down-regulated miR-144 exhibited a lower mean tumor diameter and weight, and increased apoptosis index and expression of AKAP12 and ERK1/2.

Conclusion: Taken together, these studies demonstrate the stimulative role of miR-144 in MDS progression by regulating AKAP12-dependent ERK1/2 signaling pathway.

Keywords: AKAP12; Apoptosis; ERK1/2 signaling pathway; Myelodysplastic syndrome cells; Proliferation; microRNA-144.

MeSH terms

  • A Kinase Anchor Proteins / genetics
  • A Kinase Anchor Proteins / metabolism*
  • Animals
  • Apoptosis / genetics*
  • Base Sequence
  • Cell Cycle Checkpoints / genetics
  • Cell Cycle Proteins / genetics
  • Cell Cycle Proteins / metabolism*
  • Cell Proliferation / genetics
  • Down-Regulation / genetics*
  • Female
  • Gene Expression Profiling
  • MAP Kinase Signaling System*
  • Mice, Nude
  • Mice, SCID
  • MicroRNAs / genetics
  • MicroRNAs / metabolism*
  • Models, Biological
  • Myelodysplastic Syndromes / genetics*
  • Protein Binding
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Up-Regulation / genetics

Substances

  • A Kinase Anchor Proteins
  • AKAP12 protein, human
  • Akap12 protein, mouse
  • Cell Cycle Proteins
  • MIRN144 microRNA, human
  • MIRN144 microRNA, mouse
  • MicroRNAs
  • RNA, Messenger